Two-Dimensional Electrophoresis as Applied to Problems of in Vitro and in Vivo Toxicology

Anderson, N. Leigh; Esquer-Blasco, Ricardo; Anderson, N G

doi:10.1016/b978-0-08-092440-3.50046-4

Cited by 12 publications

(11 citation statements)

References 3 publications

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“…This procedure made use of digital filtering, mathematical morphology techniques and digital masking to remove background, and used full two-dimensional least-squares optimization to refine the parameters of a 2-D Gaussian shape for each spot. The process of matching sample patterns to the master and analyzing regional protein pattern differences has been described in detail elsewhere [9]. Cortex patterns were initially matched to a cortex master pattern (KDNY-CORTEX-I, Fig.…”

Section: Gel Pattern Analysismentioning

confidence: 99%

Differential expression of cytosolic proteins in the rat kidney cortex and medulla: Preliminary proteomics

Witzmann¹,

Fultz²,

Grant³

et al. 1998

Electrophoresis

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The rodent kidney is a target of many xenobiotics and is typified by regionally specific structure and function. This renders distinct regions of the kidney differentially susceptible to toxic exposure and effect. To characterize these differences at the proteome level, protein patterns from male rat kidney cortex and medulla cytosols were examined by two-dimensional electrophoresis (2-DE) and image analysis and prominent proteins identified immunologically or by matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) and electrospray/ionization-tandem mass spectrometry (ESI-MS/MS) sequence tag identification. An average of 727 protein spots were resolved and matched to the cortex cytosol reference pattern, and 716 in the medulla. Of this total, 127 proteins were found to differ in abundance (86 higher in cortex; 41 higher in medulla) (P < 0.001). Of those proteins that were detectable in both cortex and medulla, the abundance of 97 differed significantly while 30 proteins were found to be unique to one region or the other (26 in cortex, 4 in medulla). Twenty protein spots were identified and their regional differences are discussed. These results both confirm and expand our understanding of the molecular heterogeneity characterizing structurally and functionally distinct regions of the kidney and serve as a useful foundation for future nephrotoxicologic studies.

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Section: Gel Pattern Analysismentioning

confidence: 99%

Differential expression of cytosolic proteins in the rat kidney cortex and medulla: Preliminary proteomics

Witzmann¹,

Fultz²,

Grant³

et al. 1998

Electrophoresis

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“…Stained gels were optically scanned at 121 mm resolution using a CCD scanner, and images were processed as described [24]. Groupwise statistical comparisons were made to screen for protein alterations (Student©s t-test).…”

Section: Gel Pattern Analysismentioning

confidence: 99%

Proteomic analysis of the renal effects of simulated occupational jet fuel exposure

et al. 2000

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“…The differential abundances may be attributed, once again, to the differential turnover rates of testis proteins. However, assuming accurate sample loading and, more importantly, the correction via scaling for differential loading [24] conducted on each sample, generalized protein synthesis effects are accounted for and the observed changes are related to intoxication. Again, identification of the protein spots will provide mechanistic relevance to these selective declines in abundance.…”

Section: Methodsmentioning

confidence: 99%

“…Stained gels were optically scanned at 121 micron resolution using a CCD scanner, and images were processed as described [24]. Groupwise statistical comparisons were made to screen for protein alterations (Student t-test).…”

Section: Methodsmentioning

confidence: 99%

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Protein Markers of Chemical Exposure and Molecular Epidemiology

Witzmann¹

1999

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PubKc reporting burden for this collection of information is estimated to average 1 hour per response, including the time for reviewing instructions, searching existing data sources gathering and rnaintaining the data needed, and completing and reviewing the collection of information. Send comments regarding this burden estimate or any other aspect Of this collection of information, including suggestions for reducing this burden to Washington Headquarters Services, Directorate for Information Operations and Reports, 1215 Jefferson Davis Highway, Suite 1204, Arlington, VA 222024302, and to the Office of Management and Budget, Paperwork Reduction Project (0704-0188), Washington, DC 20503. AGENCY USE ONLY (Leave Blank)2. REPORT DATE July 30,1999 TITLE AND SUBTITLE Protein Markers of Chemical Exposure and Molecular Epidemiology AUTHOR(S)Frank A. Witzmann PERFORMING ORGANIZATION NAME(S) AND ADDRESSES)Molecular Anatomy Laboratory, Dept of Biology Indiana University Purdue University -Columbus 4601 Central Avenue Columbus IN 47203 SPONSORINGyMONITORING AGENCY NAME(S) AND ADDRESS(ES)AFOSR 12b. DISTRIBUTION CODE ABSTRACT (Maximum 200 words)This final technical report describes the results of experiments that 1) explored, at the molecular level, the toxicity of lead in various exposure systems; 2) enabled the development a system of general and tissue-specific protein markers for lead in rabbits and rats that might be indicative of its effect in human toxicologic targets; 3) assessed the cellular expression of GST isoenzymes as protein markers of lead toxicity in rat kidney, and 4) investigated the suitability of tissue-slice in vitro toxicologic approaches by evaluating their patterns of protein expression and comparing them to patterns from whole tissues. High resolution large-scale two-dimensional polyacrylamide gel electrophoresis (2-DE) with computerized image analysis was used to map cell/tissue proteins of various lead targets. In the case of lead experiments, proteomics technology, e.g. mass spectrometric analysis of tryptic digests (peptide-mass fingerprinting) of selected proteins resolved by 2-DE , was used to identify altered proteins. Quantitative and qualitative assessments of toxic effect were made, individual protein data was compiled in annotated databases, and protein markers identified. Protein digestion 9 Matrix-assisted laser desorption mass spectrometric (MALDI) protein identification 9 SUBJECT TERMSResults and Discussion 1" Toxicity of lead in rabbit ventricle and kidney 10 Assessment of regional differences in renal lead toxicity 12 Assessment of glutathione S-transferase (GST) isoenzyme expression 14 AbstractThis final technical report describes the results of experiments that 1) explored, at the molecular level, the toxicity of lead in various exposure systems; 2) enabled the development a system of general and tissuespecific protein markers for lead in rabbits and rats that might be indicative of its effect in human toxicologic targets; 3) assessed the cellular expression of GST isoenzymes as ...

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Two-Dimensional Electrophoresis as Applied to Problems of in Vitro and in Vivo Toxicology

Cited by 12 publications

References 3 publications

Differential expression of cytosolic proteins in the rat kidney cortex and medulla: Preliminary proteomics

Differential expression of cytosolic proteins in the rat kidney cortex and medulla: Preliminary proteomics

Proteomic analysis of the renal effects of simulated occupational jet fuel exposure

Protein Markers of Chemical Exposure and Molecular Epidemiology

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